Methods, reagents, devices and instrumentation for preparing impregnated tissue samples suitable for histopathological and molecular studies

Abstract

A process for the production of paraffin sections of biological tissue, especially for molecular pathology studies is disclosed. In the process, the tissue sample is simultaneously fixed, dehydrated and cleared in a first step, subsequently dehydrated and cleared in a second step and infiltrated with an inert specimen matrix in a third step. The specimen can then be further embedded in a casting supporting matrix according to the standard procedures followed by any local pathology or research laboratory. A kit and a processing station for automating paraffin embedding of a tissue sample suitable for histopathological and molecular analysis is also described. A bio-indicator system is described for measuring the degree of crosslinking. A tissue sample holding means or a vial which includes a tissue sample holding means provided with a data logging device capable of registering and transmitting data regarding the sample and conditions where the sample was processed is also disclosed.

What is claimed is: 
     
       1. A vial package for a sample processing station comprising:
 a vial configured to receive and retain liquids therein, comprising a vial body and a resealable lid configured to fittingly and reversibly seal a top opening of the vial, and 
 a tissue sample cassette, 
 
       wherein the vial body, resealable lid or the tissue sample cassette is provided with a data logging device configured to electronically register and store data regarding a sample and conditions wherein said sample is processed, wherein said data logging device comprises:
 a memory storage component configured to store said data, such that said data remains in association with the vial body, resealable lid or tissue sample cassette; 
 means for monitoring time (t) of a non-preset period during which said sample is processed, which time data is stored in said memory storage component; and 
 an on/off function adapted to activate/deactivate the data logging device, such that said monitoring takes place once the data logging device has been activated, the tissue sample cassette is connected to the resealable lid by means of an elongated member 
 
       wherein the vial package is autonomous, and is configured for removable insertion into the tissue processing station. 
     
     
       2. The vial package according to  claim 1 , wherein the elongated member is configured to position the tissue sample cassette within the lower half of the inner space of the vial body when the lid seals the top opening of the vial. 
     
     
       3. The vial package according to  claim 1 , wherein said data logging device is provided on the vial body or on the resealable lid. 
     
     
       4. The vial package according to  claim 1 , wherein said data logging device is provided on the elongated member. 
     
     
       5. The vial package according to  claim 1 , wherein said data logging device is provided on the tissue sample cassette, preferably in a cavity of said tissue sample cassette, more preferably embedded in a suitable matrix in the cavity of said tissue sample cassette. 
     
     
       6. The vial package according to  claim 1 , wherein said data logging device comprises means for monitoring time (t) during which said sample is processed and means for monitoring temperature (T) of the sample in function of time (t). 
     
     
       7. The vial package according to  claim 1 , wherein said on/off function is adapted to be activated manually or automatically. 
     
     
       8. The vial package according to  claim 1 , wherein said data logging device comprises a clock and/or timer. 
     
     
       9. The vial package according to  claim 1 , wherein said data logging device comprises a thermometer. 
     
     
       10. The vial package according to  claim 1 , wherein said data logging device further comprises a sensor capable of measuring electrical conductance. 
     
     
       11. The vial package according to  claim 1 , wherein said data logging device comprises an identification means capable of identifying said sample. 
     
     
       12. The vial package according to  claim 1 , wherein said data logging device comprises a memory storage component capable of storing said registered data. 
     
     
       13. The vial package according to  claim 1 , wherein said data logging device is connectable to a reading device which is adapted to receive, read and process the data registered by said data logging device. 
     
     
       14. The vial package according to  claim 1 , wherein the vial comprises at least one breakable seal towards the top of the vial, suitable for receiving or removing fluid there through. 
     
     
       15. The vial package according to  claim 1 , wherein the vial comprises at least one breakable seal towards the top of the vial, suitable for applying positive or negative air pressure to the vial there through. 
     
     
       16. The vial package according to  claim 1 , wherein the vial comprises at least one breakable seal towards the base of the vial, suitable for receiving fluid there through. 
     
     
       17. A method for processing a tissue sample with the system of  claim 1  which comprises activating said data logging device substantially concurrently with:
 (i) the deposition of the tissue sample to be processed into the tissue sample cassette of said system, or 
 (ii) the exposure of the tissue sample cassette comprising said tissue sample to a first treatment solution comprising a fixative, in the vial of said system. 
 
     
     
       18. The method according to  claim 17 , comprising:
 (i) querying the data logging device to determine the elapsed time during which the tissue sample has been exposed to fixative, 
 (ii) comparing said elapsed time with a pre-decided cut-off value, 
 (iii) if the elapsed time is greater than said cut-off value, subjecting the tissue sample to post-fixation tissue processing steps, or, if the elapsed time is lower than said cut-off value, calculating the time needed to completely fix the tissue and continuing fixation for said time before subjecting the tissue sample to post-fixation tissue processing steps. 
 
     
     
       19. The method according to  claim 17 , comprising:
 (i) querying the data logging device to determine the integral sum of temperature as a function of time during which the tissue sample has been exposed to fixative, 
 (ii) comparing said integral sum with a pre-decided cut-off value, 
 (iii) if the integral sum is greater than said cut-off value, subjecting the tissue sample to post-fixation tissue processing steps, or, if the integral sum is lower than said cut-off value, calculating the time needed to completely fix the tissue and continuing fixation for said time before subjecting the tissue sample to post-fixation tissue processing steps. 
 
     
     
       20. The vial package according to  claim 1 , wherein the elongated member is configured to position the tissue sample cassette within the inner space of the vial, such as to immerse the tissue sample cassette in liquids to be introduced to the vial. 
     
     
       21. The vial package according to  claim 1 , wherein the physical connection between the elongated member and the tissue cassette is configured to be broken, thereby allowing processing of the sample apart from the lid. 
     
     
       22. The vial package according to  claim 1 , wherein said data logging device further comprises a means for monitoring temperature (T) of the sample in function of time (t) which temperature data is stored in said memory storage component.